CAJ | 학술논문

目的研究血液滤过联合乌司他丁治疗脓毒性休克的分子机制.方法分别取正常人血清、脓毒性休克常规治疗患者血清、脓毒性休克血液滤过联合乌司他丁治疗(CVVH-ULI)患者血清刺激人脐静脉内皮细胞,用FITC标记的白蛋白检测血管内皮细胞通透性,用罗丹明-鬼笔环肽染色检测聚合丝状肌动蛋白(F-actin)形态改变,并检测p38的磷酸化.用p38抑制剂和DMSO预处理内皮细胞,再用脓毒性休克常规治疗患者血清刺激内皮细胞,观察其对内皮细胞通透性及F-actin形态的影响.结果脓毒性休克常规治疗患者血清处理内皮细胞可使其通透性增高,F-actin重排以及p38磷酸化增加,上述变化可被CVVH-ULI所抑制.p38抑制剂可以抑制脓毒性休克常规治疗患者血清所诱导的内皮细胞通透性增高和F-actin重排.结论CVVH-ULI可通过抑制p38活化,进而抑制F-actin重排来降低脓毒性休克所诱发的血管内皮细胞高通透性.
목적 연구혈액려과연합오사타정치료농독성휴극적분자궤제.방법 분별취정상인혈청、농독성휴극상규치료환자혈청、농독성휴극혈액려과연합오사타정치료(CVVH-ULI)환자혈청자격인제정맥내피세포,용FITC표기적백단백검측혈관내피세포통투성,용라단명-귀필배태염색검측취합사상기동단백(F-actin)형태개변,병검측p38적린산화.용p38억제제화DMSO예처리내피세포,재용농독성휴극상규치료환자혈청자격내피세포,관찰기대내피세포통투성급F-actin형태적영향.결과 농독성휴극상규치료환자혈청처리내피세포가사기통투성증고,F-actin중배이급p38린산화증가,상술변화가피CVVH-ULI소억제.p38억제제가이억제농독성휴극상규치료환자혈청소유도적내피세포통투성증고화F-actin중배.결론CVVH-ULI가통과억제p38활화,진이억제F-actin중배래강저농독성휴극소유발적혈관내피세포고통투성.
Objective To investigate the molecular mechanisms of continuous venovenous hemofiltration (CVVH) combined with ulinastatin (ULI) (CVVH-ULI) for the treatment of septic shock. Methods Human umbilical endothelial cells (HUVECs) were incubated with serums isolated from normal healthy people (control), septic shock patients treated with conventional therapy (CT) or treated with CVVH combined with ULI (CVVH-ULI). Endothelial permeability was evaluated by the leakage of FITC-labeled albumin. The morphological changes of F-actin was evaluated by Rhodamine-phalloidin. The phosphorylated levels of p38 were determined by Western blot. Cells were then treated with p38inhibitor (SB203580), or DMSO, followed by incubation with serum from septic shock patients treated with conventional therapy. Endothelial permeability and F-actin rearrangements were also evaluated as noted above. Results Serum from CT group increased endothelial permeability, F-actin rearrangements, and phosphorylated levels of p38, which were inhibited by CVVH-ULI treatment. Moreover, in CT group, the serum-induced endothelial hyperpermeability and F-actin rearrangements were inhibited by SB203580, the inhibitor of p38. Conclusion CVVH combined with ulinastatin decreases endothelial hyperpermeability induced by septic shock through inhib-iting p38 MAPK pathways.